Lysis of baculovirus particles for titration

Dear all,

We like to know the titers of our virus (I know - old school, but we do !) and with the advent of qPCR for titration, this has become an increasingly simple procedure. However, one of the time and money-consuming parts is the extraction of viral DNA. We use a Qiacube for this, but it is still resource-consuming.

As some of you may have seen, Oxford Expression Technology has recently launched a titration kit, where they lyse the baculovirus particles in a lysis buffer followed by a few heating steps in a thermocycler. Obviously, you need to buy the entire kit from them (£365.00 !) and the kit will contain far more than you need. All we need is the composition of the lysis buffer or the recipe for a buffer that will lyse baculovirus particles and release the DNA upon heating.

Here comes the question - does anybody know what is in the lysis buffer ? (I have asked OET just in case their contact was asleep at the wheel, but was not surprisingly told it was a secret, which probably means it is something very simple - but what ?).

Here is a link to the protocol: http://oetltd.com/client_media/medialibrary/2012/03/baculoQUANT_all-in-one_protocol_schematic.jpg

Thanks for any input,

Svend

NB: I have put this one on private - so whatever insights you may have are only available to group members

Posted on 29-Mar-2012 23:19 CEST

Svend Kjaer

Hi Svend,
please check out the pdf that I have got from a colleague of mine and uploaded to the Files section. Hope that helps.
Best regards,
Hüseyin

Posted on 03-Apr-2012 17:44 CEST

Hüseyin Besir

Dear Hüseyin,

Thanks a lot, that was most useful - I'll have a careful read.

Best wishes,

Svend

Posted on 03-Apr-2012 20:25 CEST

Svend Kjaer

I also need Lysis buffer of baculovirus particles for titration and checking for insertion of the gene of interest.... so can anybody help me

Posted on 05-Jun-2012 18:52 CEST

peter malak

Hi,

can you please tell me where this document is uploaded to? I can't find the files section on this website.